I've come across a few publications were the authors to demonstrate phospho-CREB increase shows only the WB band of p-CREB without showing the total CREB. Is it correct ? because I always thought that to demonstrate increase in p-CREB you need to show also the total content (as it is the case for other kinases like ERK)...any suggestions? thanks!
Hi Fulvio,
you are right — total CREB should be measured as well. However, to my experience, total CREB antibody (from Cell Signaling) works not always as nice as the pCREB from the same source. Maybe this is the reason why people sometimes don't show total CREB. I guess if you have a good reason to think that total CREB levels are not very different between different samples (e.g. short treatment of cells in culture), and the increase in pCREB is strong enough, you can still rely on these results. But if, for example, you have different animals and just show pCREB values for each of them, these data will have not much meaning, right?
Hi Pavel,
thanks for you answer...I understand that in case of short time treatment it could be right to not show total CREB (although it can be also contested that also short-time treatment could increase total protein) ...but the problem is that in the paper that I read they compare different primary cultures, using only pCREB..so I'm not sure the results are reliable...I think maybe I should open a trend on PubPeer....
Well, in the case of different primary cultures, the level of total CREB can vary tremendously, of course.
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